Degertekin Bulent, Hussain Munira, Tan Jessica, Oberhelman Kelly, Lok Anna S
Division of Gastroenterology, University of Michigan Medical Center, 3912 Taubman Center, SPC 5362, Ann Arbor, MI 48109, USA.
J Hepatol. 2009 Jan;50(1):42-8. doi: 10.1016/j.jhep.2008.08.020. Epub 2008 Nov 5.
BACKGROUND/AIMS: Early detection of antiviral drug-resistant mutations enables prompt initiation of rescue therapy. The aim of this study was to determine the accuracy and sensitivity of a new line probe assay in the detection of antiviral drug-resistant HBV mutations.
One-hundred samples from 54 patients with virologic breakthrough during entecavir, lamivudine or adefovir treatment and 21 samples from 21 nucleoside-naïve patients were tested by direct sequencing and an updated line probe assay (Innogenetics, HBV DR v.3) which incorporates probes that can detect mutations at 11 positions of the reverse transcriptase region of the HBV polymerase gene.
Complete concordance between line probe and sequencing results was observed for 90/121 samples (74.3%) and 1291/1331 amino acid positions (96.9%). Testing of follow-up samples and clonal analysis of discordant samples confirmed the presence of mutations where line probe assay but not direct sequencing detected mutations. HBV DR v.3 assay consistently detected mutations present in > or = 5% of the virus population when HBV DNA concentration was > or = 4 log10copies/mL.
The updated version of the line probe assay (HBV DR v.3) has high concordance with direct sequencing in detecting antiviral drug-resistant mutations but its sensitivity in detecting mutations at some positions needs to be improved.
背景/目的：早期检测抗病毒药物耐药突变可促使及时启动挽救治疗。本研究的目的是确定一种新型线性探针检测法在检测抗病毒药物耐药性乙肝病毒突变方面的准确性和敏感性。
对54例在恩替卡韦、拉米夫定或阿德福韦治疗期间出现病毒学突破的患者的100份样本以及21例初治核苷类药物患者的21份样本进行直接测序和一种更新的线性探针检测法（Innogenetics公司，HBV DR v.3）检测，该检测法包含能检测乙肝病毒聚合酶基因逆转录酶区域11个位点突变的探针。
121份样本中的90份（74.3%）以及1331个氨基酸位点中的1291个（96.9%），线性探针检测结果与测序结果完全一致。对后续样本的检测以及对不一致样本的克隆分析证实了线性探针检测法检测到但直接测序未检测到的突变的存在。当乙肝病毒DNA浓度≥4 log10拷贝/毫升时，HBV DR v.3检测法始终能检测到病毒群体中存在比例≥5%的突变。
线性探针检测法的更新版本（HBV DR v.3）在检测抗病毒药物耐药突变方面与直接测序具有高度一致性，但其在某些位点检测突变的敏感性有待提高。
